{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,27]],"date-time":"2025-12-27T05:18:12Z","timestamp":1766812692588,"version":"3.48.0"},"reference-count":37,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,25]],"date-time":"2025-12-25T00:00:00Z","timestamp":1766620800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100004319","name":"Pfizer","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100004319","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Jos\u00e9 Carreras\/E. Donnall Thomas Endowed Chair for Cancer Research"},{"DOI":"10.13039\/100000062","name":"NIH\/NIDDK Cooperative Center of Excellence in Hematology","doi-asserted-by":"publisher","award":["U54-DK106829"],"award-info":[{"award-number":["U54-DK106829"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000050","name":"NIH\/National Heart, Lung, and Blood Institute","doi-asserted-by":"publisher","award":["T32-HL007093"],"award-info":[{"award-number":["T32-HL007093"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background\/Objectives: Approved for treatment of acute leukemia, gemtuzumab ozogamicin (GO) and inotuzumab ozogamicin (InO) are antibody\u2013drug conjugates (ADCs) that deliver a toxic calicheamicin (CLM) derivative. The resistance mechanisms to GO\/InO remain incompletely understood. Methods: We performed a genome-wide clustered regularly interspaced short palindromic repeat (CRISPR)\/Cas9 screen for CLM sensitivity genes, and then performed confirmatory cytotoxicity assays. Results: Several DNA damage pathway regulation genes were identified, most notably TP53. Across 13 acute leukemia cell lines, the six TP53-mutant cell lines (TP53MUT) were indeed 10- to 1000-fold less sensitive to CLM than the seven TP53WT cell lines. In five TP53WT\/KO syngeneic cell line pairs we generated, TP53KO cells were significantly less sensitive to CLM than their TP53WT counterparts. In TP53WT but not TP53MUT cells, the MDM2 inhibitor and p53 activator, idasanutlin, enhanced CLM cytotoxicity, demonstrating that decoupling of cells from MDM2-p53 regulation sensitizes leukemia cells to CLM. The ATM inhibitors AZD1390 and lartesertib also significantly enhanced CLM efficacy but did so independent of the TP53 status. In contrast, neither an ATR inhibitor, Chk1\/Chk2 inhibitor, Chk2 inhibitor, or a PARP inhibitor significantly impacted CLM-induced cytotoxicity across the thirteen cell lines. Together, our studies identify ATM, MDM2, and TP53\u2014which are in the same cellular response to DNA damage pathway\u2014as key modulators of CLM-induced cytotoxicity in acute leukemia cells. Conclusions: These results support further evaluation of combination therapies with corresponding small-molecule inhibitors (currently pursued for therapy of other cancers) toward clinical testing as novel strategies to increase the efficacy of CLM-based ADCs such as GO and InO.<\/jats:p>","DOI":"10.3390\/cancers18010067","type":"journal-article","created":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T02:07:58Z","timestamp":1766714878000},"page":"67","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["DNA Damage Sensing and TP53 Function as Modulators of Sensitivity to Calicheamicin-Based Antibody\u2013Drug Conjugates for Acute Leukemia"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3203-3558","authenticated-orcid":false,"given":"Camryn M.","family":"Pettenger-Willey","sequence":"first","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8580-2659","authenticated-orcid":false,"given":"George S.","family":"Laszlo","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Margery","family":"Gang","sequence":"additional","affiliation":[{"name":"Hematology\/Oncology Fellowship Program, Fred Hutchinson Cancer Center\/University of Washington, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frances M.","family":"Cole","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Colin D.","family":"Godwin","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"Erraiss","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2255-8905","authenticated-orcid":false,"given":"Pritha","family":"Chanana","sequence":"additional","affiliation":[{"name":"Shared Resources, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Allie R.","family":"Kehret","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junyang","family":"Li","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacob W.","family":"Barton","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meghann M.","family":"Yochim","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eduardo","family":"Rodr\u00edguez-Arbol\u00ed","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"},{"name":"Department of Hematology, Hospital Universitario Virgen Del Roc\u00edo, Instituto de Biomedicina de Sevilla (IBIS\/CSIC), University of Seville, 41013 Seville, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9268-3341","authenticated-orcid":false,"given":"Roland B.","family":"Walter","sequence":"additional","affiliation":[{"name":"Translational Science and Therapeutics Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA"},{"name":"Department of Medicine, Division of Hematology and Oncology, University of Washington, Seattle, WA 98195, USA"},{"name":"Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA 98195, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1038\/s41571-021-00509-w","article-title":"Towards precision medicine for AML","volume":"18","author":"Wei","year":"2021","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1182\/blood.2022016867","article-title":"Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN","volume":"140","author":"Wei","year":"2022","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1186\/s13045-023-01409-5","article-title":"The evolution of acute lymphoblastic leukemia research and therapy at MD Anderson over four decades","volume":"16","author":"Jabbour","year":"2023","journal-title":"J. Hematol. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e959","DOI":"10.1016\/S2352-3026(24)00285-0","article-title":"Frontline treatment of adults with newly diagnosed B-cell acute lymphoblastic leukaemia","volume":"11","author":"Aldoss","year":"2024","journal-title":"Lancet Haematol."},{"key":"ref_5","first-page":"46","article-title":"Acute myeloid leukemia management and research in 2025","volume":"75","author":"Kantarjian","year":"2025","journal-title":"CA Cancer J. Clin."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1001\/jamaoncol.2025.0613","article-title":"Management of adult acute lymphoblastic leukemia: A review","volume":"11","author":"Kantarjian","year":"2025","journal-title":"JAMA Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1016\/S1470-2045(14)70281-5","article-title":"Addition of gemtuzumab ozogamicin to induction chemotherapy in adult patients with acute myeloid leukaemia: A meta-analysis of individual patient data from randomised controlled trials","volume":"15","author":"Hills","year":"2014","journal-title":"Lancet Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1038\/leu.2017.187","article-title":"Gemtuzumab ozogamicin in acute myeloid leukemia","volume":"31","author":"Godwin","year":"2017","journal-title":"Leukemia"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Collados-Ros, A., Muro, M., and Legaz, I. (2024). Gemtuzumab ozogamicin in acute myeloid leukemia: Efficacy, toxicity, and resistance mechanisms-a systematic review. Biomedicines, 12.","DOI":"10.3390\/biomedicines12010208"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1182\/bloodadvances.2018026211","article-title":"Inotuzumab: From preclinical development to success in B-cell acute lymphoblastic leukemia","volume":"3","author":"Wynne","year":"2019","journal-title":"Blood Adv."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3631","DOI":"10.1002\/cncr.35505","article-title":"Inotuzumab ozogamicin in adult acute lymphoblastic leukemia: Development, current status, and future directions","volume":"130","author":"Kantarjian","year":"2024","journal-title":"Cancer"},{"key":"ref_12","first-page":"e257050","article-title":"Incorporation of immunotherapy into adult B-cell acute lymphoblastic leukemia therapy","volume":"23","author":"Haddad","year":"2025","journal-title":"J. Natl. Compr. Cancer Netw."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Rubinstein, J.D., and O\u2019Brien, M.M. (2023). Inotuzumab ozogamicin in B-cell precursor acute lymphoblastic leukemia: Efficacy, toxicity, and practical considerations. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1237738"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Collins, D.M., Bossenmaier, B., Kollmorgen, G., and Niederfellner, G. (2019). Acquired resistance to antibody-drug conjugates. Cancers, 11.","DOI":"10.3390\/cancers11030394"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1182\/blood.2024023930","article-title":"Genomic determinants of response and resistance to inotuzumab ozogamicin in B-cell ALL","volume":"144","author":"Zhao","year":"2024","journal-title":"Blood"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1182\/blood-2004-07-2784","article-title":"Influence of CD33 expression levels and ITIM-dependent internalization on gemtuzumab ozogamicin-induced cytotoxicity","volume":"105","author":"Walter","year":"2005","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1038\/nmeth.3047","article-title":"Improved vectors and genome-wide libraries for CRISPR screening","volume":"11","author":"Sanjana","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1038\/s41375-025-02571-0","article-title":"Preclinical characterization of the anti-leukemia activity of the CD123 antibody-drug conjugate, pivekimab sunirine (IMGN632)","volume":"39","author":"Cole","year":"2025","journal-title":"Leukemia"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1658","DOI":"10.1038\/s12276-022-00863-4","article-title":"DNA damage response revisited: The p53 family and its regulators provide endless cancer therapy opportunities","volume":"54","author":"Abuetabh","year":"2022","journal-title":"Exp. Mol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Zhang, H., Xu, J., Long, Y., Maimaitijiang, A., Su, Z., Li, W., and Li, J. (2024). Unraveling the guardian: P53\u2019s multifaceted role in the DNA damage response and tumor treatment strategies. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252312928"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1186\/s40364-024-00676-9","article-title":"From regulation to deregulation of p53 in hematologic malignancies: Implications for diagnosis, prognosis and therapy","volume":"12","author":"Ahmadi","year":"2024","journal-title":"Biomark. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"15026","DOI":"10.1038\/cddiscovery.2015.26","article-title":"The MDM2 small-molecule inhibitor RG7388 leads to potent tumor inhibition in p53 wild-type neuroblastoma","volume":"1","author":"Lakoma","year":"2015","journal-title":"Cell Death Discov."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"5294","DOI":"10.1038\/s41467-024-49316-8","article-title":"ATM inhibition exploits checkpoint defects and ATM-dependent double strand break repair in TP53-mutant glioblastoma","volume":"15","author":"Laverty","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Mahmoud, A.A., Raih, M.F., Sage, E.E., Ali, Q.M., Suliman, O.H., Ibrahim, S.A.E., Mohamed, O., Abdelrazeg, S., and Mohamed, S.B. (2025). The impact of mutations on TP53 protein and MicroRNA expression in HNSCC: Novel insights for diagnostic and therapeutic strategies. PLoS ONE, 20.","DOI":"10.1371\/journal.pone.0307859"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3185","DOI":"10.1200\/JCO.2011.34.8144","article-title":"Mutations and deletions of the TP53 gene predict nonresponse to treatment and poor outcome in first relapse of childhood acute lymphoblastic leukemia","volume":"29","author":"Hof","year":"2011","journal-title":"J. Clin. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1038\/leu.2016.263","article-title":"The impact of TP53 mutations and TP53 deletions on survival varies between AML, ALL, MDS and CLL: An analysis of 3307 cases","volume":"31","author":"Stengel","year":"2017","journal-title":"Leukemia"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1038\/s41408-024-01186-5","article-title":"What have we learned about TP53-mutated acute myeloid leukemia?","volume":"14","author":"Shahzad","year":"2024","journal-title":"Blood Cancer J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1002\/ajh.27655","article-title":"TP53-mutated myeloid neoplasms: 2024 update on diagnosis, risk-stratification, and management","volume":"100","author":"Shah","year":"2025","journal-title":"Am. J. Hematol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0165-7992(90)90046-M","article-title":"Sensitivity of fibroblasts derived from ataxia-telangiectasia patients to calicheamicin gamma 1I","volume":"245","author":"Sullivan","year":"1990","journal-title":"Mutat. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"90","DOI":"10.3892\/or.2024.8749","article-title":"Combining inotuzumab ozogamicin with PARP inhibitors olaparib and talazoparib exerts synergistic cytotoxicity in acute lymphoblastic leukemia by inhibiting DNA strand break repair","volume":"52","author":"Ida","year":"2024","journal-title":"Oncol. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1186\/s12967-024-05838-9","article-title":"Exploring the role of PARP1 inhibition in enhancing antibody-drug conjugate therapy for acute leukemias: Insights from DNA damage response pathway interactions","volume":"22","author":"Jandoubi","year":"2024","journal-title":"J. Transl. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1158\/1078-0432.CCR-21-1295","article-title":"Results from a first-in-human phase I study of siremadlin (HDM201) in patients with advanced wild-type TP53 solid tumors and acute leukemia","volume":"28","author":"Stein","year":"2022","journal-title":"Clin. Cancer Res."},{"key":"ref_33","first-page":"4147","article-title":"Idasanutlin plus cytarabine in relapsed or refractory acute myeloid leukemia: Results of the MIRROS trial","volume":"6","author":"Konopleva","year":"2022","journal-title":"Blood Adv."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1182\/blood.2022016362","article-title":"Venetoclax and idasanutlin in relapsed\/refractory AML: A nonrandomized, open-label phase 1b trial","volume":"141","author":"Daver","year":"2023","journal-title":"Blood"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e70028","DOI":"10.1002\/cam4.70028","article-title":"Phase 1 dose escalation study of the MDM2 inhibitor milademetan as monotherapy and in combination with azacitidine in patients with myeloid malignancies","volume":"13","author":"DiNardo","year":"2024","journal-title":"Cancer Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4429","DOI":"10.1158\/1078-0432.CCR-25-1648","article-title":"A first-in-human study of ATM inhibitor lartesertib as monotherapy in patients with advanced solid tumors","volume":"31","author":"Siu","year":"2025","journal-title":"Clin. Cancer Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Nishikawa, S., and Iwakuma, T. (2023). Drugs targeting p53 mutations with FDA approval and in clinical trials. Cancers, 15.","DOI":"10.3390\/cancers15020429"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/1\/67\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,27]],"date-time":"2025-12-27T05:16:21Z","timestamp":1766812581000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/18\/1\/67"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,25]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["cancers18010067"],"URL":"https:\/\/doi.org\/10.3390\/cancers18010067","relation":{},"ISSN":["2072-6694"],"issn-type":[{"type":"electronic","value":"2072-6694"}],"subject":[],"published":{"date-parts":[[2025,12,25]]}}}